Extracellular vesicles (EVs) have been increasingly recognized as carriers of messages in cell-cell communication and biomarkers for different diseases, as well as for gene and drug delivery (Lee et al., Hum Mol Genet 21(R1):R125-34, 2012). EVs can be formed internally by invagination of the plasma membrane into endosomes, then in-budding of vesicles into endosomal-derived multivesicular bodies, and later fusion with the plasma membrane to release EVs into the intercellular surrounding (Thery et al., Nat Rev Immunol 9:581-593, 2009; Camussi et al., Kidney Int 78:838-848, 2010; Cocucci et al., Trends Cell Biol 19:43-51, 2009). EVs are also formed and released directly from the plasma membrane during cytoskeletal rearrangement, blebbing, or apoptosis (Camussi et al., Kidney Int 78:838-848, 2010). Cancer cells may also release a subpopulation of retroviral-like particles that are likely generated upon increased transcription of endogenous retroviral sequences (Contreras-Galindo et al., J Virol 82:9329-9336, 2008; Balaj et al., Nat Commun 2:180, 2011). Isolation and purification of released EVs remains a challenge. Methods currently used include differential and high speed ultracentrifugation (UC), separation on density gradients (Thery et al., Curr Protoc Cell Biol Chapter 3, Unit 3.22, 2006), proprietary commercial kits (e.g., EXOQUICK-TC™ kits), immune-affinity purification (Taylor et al., Methods Mol Biol 728:235-246, 2011; Tauro et al., Methods 56:293-304, 2012), and microfluidics (Chen et al., Lab Chip 10:505-511, 2010). UC, in addition to requiring specialized and expensive equipment, allows sedimentation of different types of EVs, including large oncosomes (Di Vizio et al., Cancer Res 69:5601-5609, 2009) and apoptotic bodies (Camussi et al., Kidney Int 78:838-848, 2010; Al-Nedawi et al., Cell Cycle 8:2014-2018, 2009) along with co-sedimentation of protein aggregates, such as BSA (Webber et al., J Extracell Vesicles doi: 10.3402/jev.v2i0.19861, 2013) and nucleic acids (Muller et al., J Immunol Methods 411C:55-65, 2014). Furthermore, EVs tend to cluster together and to form large aggregates in the pellet which are difficult to separate and may alter uptake of EVs by recipient cells (Momen-Heravi et al., Front Physiol 3:162, 2012). Density gradients are lengthy and laborious with low yield, and density may not be the best criteria to separate EVs, as it may vary significantly between samples. Other methods do not allow large scale EV isolation and/or require cocktails of disease-specific antibodies as well as lengthy optimizations. Therefore, there is a need for improved methods of efficiently isolating EVs.